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Accurate quantification of gene and transcript-specific expression, with the underlying knowledge of precise transcript isoforms, is crucial to understanding many biological processes. Analysis of RNA sequencing data has benefited from the development of alignment-free algorithms which enhance the precision and speed of expression analysis. However, such algorithms require a reference transcriptome. Here we generate a reference transcript dataset (LsRTDv1) for lettuce (cv. Saladin), combining long- and short-read sequencing with publicly available transcriptome annotations, and filtering to keep only transcripts with high-confidence splice junctions and transcriptional start and end sites. LsRTDv1 identifies novel genes (mostly long non-coding RNAs) and increases the number of transcript isoforms per gene in the lettuce genome from 1.4 to 2.7. We show that LsRTDv1 significantly increases the mapping rate of RNA-seq data from a lettuce time-series experiment (mock- and Botrytis cinerea-inoculated) and enables detection of genes that are differentially alternatively spliced in response to infection as well as transcript-specific expression changes. LsRTDv1 is a valuable resource for investigation of transcriptional and alternative splicing regulation in lettuce.
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Diabetes mellitus (DM) is a prevalent metabolic disorder often accompanied by oxidative stress, which contributes to various diabetic complications. Investigating the antioxidant activity of linalool (LIN) is crucial as it may offer a natural therapeutic approach to mitigate oxidative damage in DM. The aim of the present study was to investigate the antioxidant activity of LIN in a DM rat model. A total of 40 male Wistar albino rats (age, 8 weeks; weight, 250-300 g) were used. CONTROL and DM groups were administered physiological saline solution by oral gavage for 21 days. In rats in the DM + LIN and LIN groups, 100 mg/kg LIN was administered intragastrically after streptozotocin injection (n=10 per group). In the first (48 h after STZ injection), second (1 week later), third (2 weeks later), and fourth (3 weeks later) blood glucose measurements, a statistically significant increase was found in the blood glucose values of the DM and DM + LIN groups compared with those of the CONTROL group. During the 21-day experimental period, there was no reduction in blood glucose levels of the DM + LIN group. Consequently, no discernible anti-hyperglycemic effect of LIN was observed. Catalase enzyme activity, superoxide dismutase (SOD) enzyme activity, malondialdehyde (MDA) levels and glutathione (GSH) levels were measured spectrophotometrically. All assays were conducted according to the protocols provided in the respective kits. The results were analyzed to assess the oxidative status and antioxidant capacity in the experimental groups. Catalase (CAT) activity was decreased in the DM group compared with that in the CONTROL group in both the serum and liver. However, LIN administration restored CAT activity in the DM + LIN group to the level of the CONTROL group. In the liver, the DM + LIN-treated group showed a notable reduction in malondialdehyde (MDA) levels compared with those in the DM group. In conclusion, the present results suggest that the antioxidant properties of LIN may have a regulatory effect on the oxidative status in diabetes-affected systems, potentially offering therapeutic benefits in managing oxidative stress associated with diabetes.
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Healthy nutrition is widely considered the cornerstone of optimal athletic performance, both physically and mentally. This study investigates the critical role of healthy nutrition in shaping the physical and mental performance of athletes, with a specific focus on taekwondo. This research aims to explore the potential relationship between taekwondo athletes' attitudes towards healthy nutrition and their mental toughness. The research group comprised 276 active and licensed taekwondo athletes who voluntarily participated in the study (Age M = 17.18 ± SD = 7.13, N = 125 women, N = 151 men). Ethical approval for the research was obtained prior to the commencement of the study. Data collection instruments included the Healthy Nutrition Attitude Scale, Mental Toughness Scale, and a personal information form. Confirmatory factor analysis was conducted to confirm the validity and reliability of the scales. Descriptive statistics, correlation analysis, and regression analysis were performed to explore the relationship between the variables within the research model. The correlation analysis identified two significant, positive, and moderate correlations: (1) between knowledge about nutrition and mental toughness (r = 0.626) and (2) between positive nutrition and mental toughness (r = 0.672). The regression analysis revealed that both knowledge about nutrition (ß = 0.360) and positive nutrition (ß = 0.461) significantly contribute to mental toughness. The findings suggest that as athletes' knowledge of nutrition expands and their attitudes towards healthy eating become more positive, their mental toughness also appears to improve. These results are both important and original, adding significant new insights to the existing research landscape.
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Rendimiento Atlético , Dieta Saludable , Masculino , Humanos , Femenino , Reproducibilidad de los Resultados , AtletasRESUMEN
BACKGROUND: This study aimed to analyze the predictive effect of various inflammatory indices and inflammatory biomarkers on prognosis after coronary artery bypass grafting (CABG). METHODS: In this retrospective observational study, data were recorded from 99 patients who underwent isolated elective CABG between January 2019 and June 2021 and met the inclusion criteria. The patients were divided into two main groups according to the postoperative clinical results: "favorable outcome" and "poor outcome." Preoperative inflammatory parameters, inflammatory indices (such as systemic inflammation index (SII), neutrophil/lymphocyte ratio (NLR), derived NLR (dNLR)), and clinical variables were compared between the groups. RESULTS: Poor postoperative outcomes developed in 31 (31.3%) patients. In the univariate analysis, white blood cell count (p=0.008), neutrophil count (p=0.002), SII (p=0.018), NLR (p=0.003), and dNLR (p=0.003) were found to be significant predictors for poor outcomes. In the multivariate analysis, only the presence of chronic obstructive pulmonary disease (COPD) (OR=8.765; 95% CI 1.308-58.702; p=0.025) and high creatinine levels (OR=1.049; 95% CI 1.005-1.094; p=0.027) were independent risk factors for poor outcomes. Optimal cut-off values were 603.08 (areas under the curve (AUC)=0.632, p=0.036) for SII, 2.34 (AUC=0.669, p=0.007) for NLR, and 1.76 (AUC=0.667, p=0.008) for dNLR. CONCLUSION: SII, NLR, dNLR, and inflammatory markers, such as white blood cell and neutrophil counts, are feasible markers for predicting poor outcomes following CABG procedures. These parameters may aid in the development of early therapeutic interventions to improve patient outcomes.
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Muscle injury is a common type of soft tissue injury. Increased oxidative damage has been reported after muscle injuries. Therapeutic ultrasound is commonly used for such injuries. This study compared the efficacy of therapeutic ultrasound treatment and various antioxidant agents in experimental muscle injuries. For this purpose, some serum enzymes, oxidative stress, and inflammatory markers were evaluated together with histopathological examinations. Six groups were formed with 6 male Wistar albino rats in each group. These groups were control, only injury (OI), ultrasound (U), vitamin C (Vit C), selenium (S), and mixture (M). Muscle injury was caused by a laceration of the gastrocnemius muscle in all groups except the control group. No treatment was performed in the OI group. At the end of the 6-day application, all rats were sacrificed. As for serum enzymes, CK, ALT, and AST levels returned to control values in almost all treatment groups. Total oxidative status (TOS) and oxidative stress index (OSI) increased in the OI group, while they decreased in the S and M groups. In addition, the decrease in MPO activity in the blood tissue of the Vit C group was statistically significant. There were no significant changes between groups in terms of serum inflammatory markers and histological findings. This study has shown that the ingestion of vitamin C and selenium may contribute to the treatment of muscle injury in addition to therapeutic ultrasound treatment. However, further studies are needed to support these results.
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Selenio , Terapia por Ultrasonido , Ratas , Masculino , Animales , Ácido Ascórbico/farmacología , Ácido Ascórbico/uso terapéutico , Selenio/farmacología , Ratas Wistar , Antioxidantes/farmacología , Estrés Oxidativo , Vitaminas/farmacología , Músculo EsqueléticoRESUMEN
OBJECTIVE: The aim of this study was to determine the levels of resolvin D1 (RVD1) in the gingival crevicular fluid (GCF) and saliva in the patients with periodontitis and healthy subjects, and also to evaluate the effects of non-surgical periodontal treatments (NSPTs) on RVD1 levels. MATERIALS AND METHODS: Fifteen patients with Stage III Grade B periodontitis (P) and 11 periodontally healthy individuals (H) were included in this study. Clinical periodontal measurements, GCF, and saliva samples were collected from each individual at baseline and 6 weeks after NSPTs in periodontitis group. GCF and saliva levels of RVD1 were analyzed by enzyme-linked immunosorbent assay. RESULTS: GCF total and concentration levels of RVD1 were significantly lower in the periodontitis group than in the healthy group and significantly increased after NSPTs in periodontitis (p < 0.001). There was no statistically significant difference in saliva RVD1 levels between healthy and periodontitis group and also before and after NSPTs in periodontitis (p > 0.05). Significant negative correlations were found between all periodontal clinical parameters and GCF volume with both GCF total amount and concentrations of RVD1 (p < 0.01). There was a positive correlation between GCF total amount and concentrations of RVD1 (r = 0.762, p < 0.01). CONCLUSIONS: GCF levels of RVD1 might be promising biomarkers for monitoring the susceptibility to periodontitis and predicting periodontal status. CLINICAL RELEVANCE: RVD1 may be valuable biomarker to observe the healing process after periodontal treatment as increased GCF levels might project clinical improvements post-treatment. Accordingly, observing GCF RVD1 levels might be helpful to determine individuals require further periodontal treatment.
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Periodontitis Crónica , Biomarcadores/análisis , Periodontitis Crónica/terapia , Ácidos Docosahexaenoicos , Líquido del Surco Gingival/química , HumanosRESUMEN
The oncogenic gammaherpesviruses, including human Epstein-Barr virus (EBV), human Kaposi's sarcoma-associated herpesvirus (KSHV), and murine gammaherpesvirus 68 (MHV68, γHV68, MuHV-4), are associated with numerous malignancies, including B cell lymphomas and nasopharyngeal carcinoma. These viruses employ numerous molecular strategies to colonize the host, including the expression of noncoding RNAs (ncRNAs). As the first viral ncRNAs identified, EBV-encoded RNA 1 and 2 (EBER1 and EBER2, respectively) have been investigated extensively for decades; however, their specific in vivo functions remain largely unknown. In work here, we used chimeric MHV68 viruses in an in vivo complementation system to test whether EBV EBER2 contributes to acute and/or chronic phases of infection. Expression of EBER2 derived from EBV strain B95-8 resulted in a significant expansion of latently infected B cells in vivo, which was accompanied by a decrease in virus-infected plasma cells. EBV strains typically carry one of two variants of EBER2, which differ primarily by a 5-nucleotide core polymorphism identified initially in the EBV strain M81. Strikingly, mutation of the 5 nucleotides that define this core polymorphism resulted in the loss of the infected B cell expansion and restored plasma cell infection. This work reveals that the B95-8 variant of EBER2 promotes the expansion of the latently infected B cell pool in vivo and may do so in part through inhibition of terminal differentiation. These findings provide new insight into mechanisms by which viral ncRNAs promote in vivo colonization and further and provide further evidence of the inherent tumorigenic risks associated with gammaherpesvirus manipulation of B cell differentiation. IMPORTANCE The oncogenic gammaherpesviruses, including human Epstein-Barr virus (EBV), human Kaposi's sarcoma-associated herpesvirus (KSHV), and murine gammaherpesvirus 68, employ numerous strategies to colonize the host, including expression of noncoding RNAs (ncRNAs). As the first viral ncRNAs ever identified, EBV-encoded RNA 1 and 2 (EBER1 and EBER2) have been investigated extensively for decades; however, their specific in vivo functions remain largely unknown. Work here reveals that an EBV EBER2 variant highly associated with B cell lymphoma promoted a significantly increased expansion of the infected B cell pool in vivo, which coincided with altered B cell differentiation. Mutation of the 5 nucleotides that define this EBER2 variant resulted in the loss of B cell expansion and normal B cell differentiation. These findings provide new insight into the mechanisms by which EBV manipulates B cells in vivo to retain infected cells in the high-risk B cell differentiation pathway where they are poised for tumorigenesis.
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Infecciones por Virus de Epstein-Barr , Gammaherpesvirinae , Herpesvirus Humano 8 , Rhadinovirus , Animales , Infecciones por Virus de Epstein-Barr/genética , Gammaherpesvirinae/genética , Herpesvirus Humano 4/fisiología , Herpesvirus Humano 8/genética , Humanos , Ratones , Nucleótidos , Polimorfismo Genético , ARN no Traducido/genética , ARN no Traducido/metabolismo , ARN Viral , Rhadinovirus/genética , Latencia del Virus/genéticaRESUMEN
Cluster randomized trials are frequently used in educational research for methodological reasons. This study aims to improve the efficiency of cluster randomized trials on computer/information literacy and computational thinking. The study employs a two-level hierarchical linear model to estimate (i) intraclass correlation coefficients, (ii) the amount of explained variances given selected predictors, and (iii) minimum detectable effect sizes given the set of plausible scenarios. Two data cycles from the International Computer and Information Study were used. The covariates at the student level are gender, interest in ICT, parents' highest education level, ICT self-efficacy, and experience with computers. The covariates at school/teacher level are teacher's ICT use, ratio of school size to the number of computers for student use, availability of ICT resources at school, approximate teacher age, and ICT self-efficacy. Findings showed that the most precise effect could be measured when student and teacher/school covariates are both adopted. Lastly, it was revealed that increasing the number of schools is effective to get the most precise effect.
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BACKGROUND: Respiratory failure and death are the leading causes of severe Coronavirus disease 2019 (COVID-19). Hyper-inflammation and cytokine storm cause lung damage. This study aimed to compare the low-dose and high-dose effects of tocilizumab, an IL-6 receptor antagonist. METHOD: Patients with severe pneumonia and hyper-inflammation signs because of COVID-19 were included in this retrospective study. Patients receiving tocilizumab <200 mg intravenously were classified as the low-dose group, and receiving ≥200 mg as the high-dose group, and those not treated with tocilizumab as the control group. Demographic and clinical data of patients who died and survived in both low-high dose and control patients were compared. According to symptom day and radiological infiltration, patients with tocilizumab were also evaluated in two groups as early and late periods at tocilizumab administration time. RESULTS: A total of 160 patients were included in the study; 70 were treated with a low dose and 50 with high-dose tocilizumab. Forty patients were in the control group. Age, comorbidity and clinical features were similar in the control, low-dose tocilizumab and high-dose tocilizumab groups. The mortality rate (12.9%, 30.0%, 37.5, P = .008) was less in the low-dose tocilizumab group. The secondary infection rate was higher in the high-dose group than in the low-dose tocilizumab and control groups (44.0%, 10.0%, 10.0%, P < .001). Distinguishing between those patients who died and survived, age (OR: 1.1589, P < .001), higher APACHE II scores (OR: 1.225, P = .001) and needs for non-invasive mechanical ventilation (OR: 14.469, P < .001) were the most critical risk factors. Low-dose tocilizumab was associated with a lower mortality rate (OR: 0.244, P = .012). CONCLUSION: The use of tocilizumab at a low dose is associated with lower secondary infections and mortality.
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Anticuerpos Monoclonales Humanizados/uso terapéutico , Tratamiento Farmacológico de COVID-19 , Coinfección , Coinfección/prevención & control , Humanos , Estudios Retrospectivos , Resultado del TratamientoRESUMEN
OBJECTIVE: Epilepsy is a common disorder that affects the nervous systems of 1% of worldwide population. In epilepsy, one-third of patients are unresponsive to current drug therapies and develop drug-resistant epilepsy. Alterations in ghrelin, nesfatin-1, and irisin levels with epilepsy were reported in previous studies. Vasoactive intestinal peptide is among the most common neuropeptides in the hippocampus, which is the focus of the seizures in temporal lobe epilepsy. However, there is also lack of evidence of whether these four neuropeptide levels are altered with drug resistant temporal lobe epilepsy or not. The aim herein was the evaluation of the serum levels of nesfatin-1, ghrelin, irisin, and Vasoactive intestinal peptide in drug-resistant temporal lobe epilepsy patients and temporal lobe epilepsy (TLE) without drug resistance, and to compare them to healthy controls. METHODS: This cross-sectional study group included 58 temporal lobe epilepsy patients (24 with drug resistant temporal lobe epilepsy and 34 with temporal lobe epilepsy who were not drug-resistant) and 28 healthy subjects. Nesfatin-1, ghrelin, irisin, and Vasoactive intestinal peptide serum levels were determined using enzyme-linked immunosorbent assay. RESULTS: The serum ghrelin levels of patients with drug resistant temporal lobe epilepsy were seen to have significantly decreased when compared to those of the control group (p<0.05). Serum nesfatin-1, vasoactive intestinal peptide, and irisin levels were seen to have decreased in the drug resistant temporal lobe epilepsy group when compared to those of the control and temporal lobe epilepsy groups; however, the difference was non-significant (p>0.05). CONCLUSIONS: The results herein suggested that ghrelin might contribute to the pathophysiology of drug resistant temporal lobe epilepsy. However, further studies are needed to confirm this hypothesis.
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Epilepsia del Lóbulo Temporal , Fibronectinas , Ghrelina , Nucleobindinas , Péptido Intestinal Vasoactivo , Estudios Transversales , Resistencia a Medicamentos , Epilepsia del Lóbulo Temporal/tratamiento farmacológico , HumanosRESUMEN
Viruses, like their metazoan hosts, have evolved to utilize intricate transcriptional mechanisms to generate a vast array of both coding and noncoding RNA transcripts. The resolution of specific noncoding RNA transcripts produced by viruses, particularly herpesviruses, presents a particularly difficult challenge due to their highly dense dsDNA genomes and their complex, overlapping, and context-dependent network of transcripts. While new long read sequencing platforms have facilitated the resolution of some noncoding transcripts from virus genomes, empirical molecular validation of transcripts from individual regions is essential. Herein, we demonstrate that the use of strand specific northern blots is essential for true validation of specific viral noncoding RNAs, and provide here a detailed molecular method for such an approach.
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Northern Blotting , Genes Sobrepuestos , ARN Largo no Codificante/genética , ARN Mensajero/genética , ARN Viral/genética , Northern Blotting/métodos , Electroforesis en Gel de Poliacrilamida , Regulación Viral de la Expresión Génica , Genoma Viral , Herpesviridae/genética , Sistemas de Lectura Abierta , Virus/genéticaRESUMEN
The Epstein Barr virus (EBV) contributes to the tumor phenotype through a limited set of primarily non-coding viral RNAs, including 31 mature miRNAs. Here we investigated the impact of EBV miRNAs on remodeling the tumor cell transcriptome. Strikingly, EBV miRNAs displayed exceptionally abundant expression in primary EBV-associated Burkitt's Lymphomas (BLs) and Gastric Carcinomas (GCs). To investigate viral miRNA targeting, we used the high-resolution approach, CLASH in GC and BL cell models. Affinity constant calculations of targeting efficacies for CLASH hits showed that viral miRNAs bind their targets more effectively than their host counterparts, as did Kaposi's sarcoma-associated herpesvirus (KSHV) and murine gammaherpesvirus 68 (MHV68) miRNAs. Using public BL and GC RNA-seq datasets, we found that high EBV miRNA targeting efficacies translates to enhanced reduction of target expression. Pathway analysis of high efficacy EBV miRNA targets showed enrichment for innate and adaptive immune responses. Inhibition of the immune response by EBV miRNAs was functionally validated in vivo through the finding of inverse correlations between EBV miRNAs and immune cell infiltration and T-cell diversity in BL and GC datasets. Together, this study demonstrates that EBV miRNAs are potent effectors of the tumor transcriptome that play a role in suppressing host immune response.
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Infecciones por Virus de Epstein-Barr/inmunología , Regulación Viral de la Expresión Génica , Herpesvirus Humano 4/inmunología , MicroARNs/genética , ARN Viral/genética , Complejo Silenciador Inducido por ARN/metabolismo , Transcriptoma , Infecciones por Virus de Epstein-Barr/metabolismo , Infecciones por Virus de Epstein-Barr/patología , Infecciones por Virus de Epstein-Barr/virología , Herpesvirus Humano 4/genética , Humanos , Complejo Silenciador Inducido por ARN/genéticaRESUMEN
SUMMARY OBJECTIVE: Epilepsy is a common disorder that affects the nervous systems of 1% of worldwide population. In epilepsy, one-third of patients are unresponsive to current drug therapies and develop drug-resistant epilepsy. Alterations in ghrelin, nesfatin-1, and irisin levels with epilepsy were reported in previous studies. Vasoactive intestinal peptide is among the most common neuropeptides in the hippocampus, which is the focus of the seizures in temporal lobe epilepsy. However, there is also lack of evidence of whether these four neuropeptide levels are altered with drug resistant temporal lobe epilepsy or not. The aim herein was the evaluation of the serum levels of nesfatin-1, ghrelin, irisin, and Vasoactive intestinal peptide in drug-resistant temporal lobe epilepsy patients and temporal lobe epilepsy (TLE) without drug resistance, and to compare them to healthy controls. METHODS: This cross-sectional study group included 58 temporal lobe epilepsy patients (24 with drug resistant temporal lobe epilepsy and 34 with temporal lobe epilepsy who were not drug-resistant) and 28 healthy subjects. Nesfatin-1, ghrelin, irisin, and Vasoactive intestinal peptide serum levels were determined using enzyme-linked immunosorbent assay. RESULTS: The serum ghrelin levels of patients with drug resistant temporal lobe epilepsy were seen to have significantly decreased when compared to those of the control group (p<0.05). Serum nesfatin-1, vasoactive intestinal peptide, and irisin levels were seen to have decreased in the drug resistant temporal lobe epilepsy group when compared to those of the control and temporal lobe epilepsy groups; however, the difference was non-significant (p>0.05). CONCLUSIONS: The results herein suggested that ghrelin might contribute to the pathophysiology of drug resistant temporal lobe epilepsy. However, further studies are needed to confirm this hypothesis.
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Humanos , Péptido Intestinal Vasoactivo , Fibronectinas , Epilepsia del Lóbulo Temporal/tratamiento farmacológico , Ghrelina , Nucleobindinas , Resistencia a Medicamentos , Estudios TransversalesRESUMEN
OBJECTIVE: This study aimed to evaluate the effects of different surface conditoning methods on surface texture and shear bond strength (SBS) of brackets bonded to resin nanoceramic material. METHODS: Ceramic specimens were divided into two groups as metal brackets and ceramic brackets. In each group, the following five subgroups were conditoned with orthophosphoric acid (OPA), hydrofluoric acid (HFA), silica coating with Cojet, Nd: Yag laser, and Femtosecond (Fs) laser. Extra samples were used for scanning electron microscopy and 3D profilometer evaluation. RESULTS: All surface conditioning methods caused optimum or higher SBS. Metal brackets had higher SBS than porcelain brackets, but this difference reached statistical significance only in Fs laser group. OPA caused surface modification comparable to HFA because of polymer content of resin nanoceramic. Although Fs laser and Cojet conditioning caused optimum or higher SBS, surface damage of these methods to the resin nanoceramic specimens clearly seen on 3D profilometer. CONCLUSION: HFA and Nd: Yag laser are effective surface conditioning methods for resin nanoceramics. OPA combined with silane application caused optimum SBS and can be used as an alternative to HFA. Surface texture changes should be considered to determine surface damage while deciding the optimum surface conditioning method for ceramics other than SBS.
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Today, environmental problems are increasing and threatening nature and human health. Air pollution is at the top of this threat. Air, the main source of life, is indispensable for humans and living things. Therefore, air pollution causes mass results. Many air pollution studies and many solution techniques have been proposed in the literature to deal with the air pollution problem. In this study, it is called analytical hierarchy process (AHP) and geographic information systems (GIS), which is one of the multi-criteria decision making methods used in the investigation of air pollution in Igdir city center and its four districts: Tuzluca, Igdir Central, Karakoyunlu, and Aralik. In this study, spatial analysis of the pollutant parameters using the GIS-AHP technique was performed with the help of the data obtained from Igdir Weather Monitoring Stations. By determining the pollutant parameters, pollution distribution maps were created, and station-based statistics were evaluated with dynamic mapping. For the first time, 15 parameters of weather and topographic features were used. Based on this result, it is obvious that Igdir center is the worst place in the air pollution problem for GIS-AHP method. It varies for 15 parameters using GIS-AHP, compared with the review of locations on a daily basis. The aim of this study is primarily to investigate possible places of air pollution. Also, by comparing the results of two methods, GIS and AHP, more accurate results are given.
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Contaminantes Atmosféricos/análisis , Contaminación del Aire/análisis , Ciudades , Monitoreo del Ambiente , Sistemas de Información Geográfica , Humanos , Material Particulado/análisis , TurquíaRESUMEN
OBJECTIVES: We evaluated whether differences exist among the canal flare index (CFI) values obtained by different calculation methods in the veterinary literature. STUDY DESIGN: The endosteal widths were measured from radiographic images of canine cadaveric femora. Three different formulae were used to calculate the CFI. The CFILT-M was the ratio of endosteal width at the medial aspect of the lesser trochanter (LT) to the midshaft (M), while the CFILT-I was the ratio of LT to the isthmus (I). The CFIPLT-I was the ratio of endosteal width at the proximal aspect of the lesser trochanter (PLT) and the I. The widths at each level and the CFI calculation methods were compared. Using Rashmir-Raven's method, the femora were typed as stovepipe, normal and champagne fluted. The limits of agreement were also evaluated. RESULTS: The endosteal width at the proximal aspect of the lesser trochanter was 12% wider than at the medial aspect and 8% wider at the midshaft than at the isthmus. The CFILT-M was less than CFILT-I and CFIPLT-I by 9 and 20%, respectively. By Rashmir-Raven's classification, the CFILT-M method provided 18% stovepipe, 79% normal and 3% champagne fluted femora. The CFILT-I method showed the stovepipe, the normal and the champagne fluted as 6, 82 and 12%, respectively. The CFIPLT-I method classified the femora either normal (55%) or champagne fluted (45%). The comparison of CFILT-M with the other methods using Bland-Altman analysis showed lower mean difference for the CFILT-I than the CFIPLT-I. CONCLUSION: The level of width measurements at proximal femora might have an impact on the CFI values, likewise, preoperative planning procedures and the selection of a stem type in total hip arthroplasty.
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Perros/anatomía & histología , Fémur/anatomía & histología , Animales , Pesos y Medidas Corporales/métodos , Pesos y Medidas Corporales/veterinaria , Cadáver , Femenino , Fémur/diagnóstico por imagen , MasculinoRESUMEN
Gammaherpesviruses, including the human pathogens Epstein-Barr virus (EBV) and Kaposi's sarcoma-associated herpesvirus (KSHV), establish lifelong latent infection in B cells and are associated with a variety of tumors. In addition to protein coding genes, these viruses encode numerous microRNAs (miRNAs) within their genomes. While putative host targets of EBV and KSHV miRNAs have been previously identified, the specific functions of these miRNAs during in vivo infection are largely unknown. Murine gammaherpesvirus 68 (MHV68) is a natural pathogen of rodents that is genetically related to both EBV and KSHV, and thus serves as an excellent model for the study of EBV and KSHV genetic elements such as miRNAs in the context of infection and disease. However, the specific targets of MHV68 miRNAs remain completely unknown. Using a technique known as qCLASH (quick crosslinking, ligation, and sequencing of hybrids), we have now identified thousands of Ago-associated, direct miRNA-mRNA interactions during lytic infection, latent infection and reactivation from latency. Validating this approach, detailed molecular analyses of specific interactions demonstrated repression of numerous host mRNA targets of MHV68 miRNAs, including Arid1a, Ctsl, Ifitm3 and Phc3. Notably, of the 1,505 MHV68 miRNA-host mRNA targets identified in B cells, 86% were shared with either EBV or KSHV, and 64% were shared among all three viruses, demonstrating significant conservation of gammaherpesvirus miRNA targeting. Pathway analysis of MHV68 miRNA targets further revealed enrichment of cellular pathways involved in protein synthesis and protein modification, including eIF2 Signaling, mTOR signaling and protein ubiquitination, pathways also enriched for targets of EBV and KSHV miRNAs. These findings provide substantial new information about specific targets of MHV68 miRNAs and shed important light on likely conserved functions of gammaherpesvirus miRNAs.
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Gammaherpesvirinae/fisiología , Infecciones por Herpesviridae/metabolismo , MicroARNs/genética , Procesamiento Proteico-Postraduccional , ARN Mensajero/metabolismo , Serina-Treonina Quinasas TOR/metabolismo , Factores de Transcripción/metabolismo , Animales , Regulación de la Expresión Génica , Infecciones por Herpesviridae/genética , Infecciones por Herpesviridae/virología , Ratones , ARN Mensajero/genética , ARN Viral/genética , ARN Viral/metabolismo , Serina-Treonina Quinasas TOR/genética , Factores de Transcripción/genética , Replicación ViralRESUMEN
The gammaherpesviruses, including Epstein-Barr virus (EBV), Kaposi's sarcoma-associated herpesvirus (KSHV), and murine gammaherpesvirus 68 (MHV68, MuHV-4, γHV68), are etiologic agents of a wide range of lymphomas and non-hematological malignancies. These viruses possess large and highly dense dsDNA genomes that feature >80 bidirectionally positioned open reading frames (ORFs). The abundance of overlapping transcripts and extensive splicing throughout these genomes have until now prohibited high throughput-based resolution of transcript structures. Here, we integrate the capabilities of long-read sequencing with the accuracy of short-read platforms to globally resolve MHV68 transcript structures using the transcript resolution through integration of multi-platform data (TRIMD) pipeline. This approach reveals highly complex features, including: (1) pervasive overlapping transcript structures; (2) transcripts containing intra-gene or trans-gene splices that yield chimeric ORFs; (3) antisense and intergenic transcripts containing ORFs; and (4) noncoding transcripts. This work sheds light on the underappreciated complexity of gammaherpesvirus transcription and provides an extensively revised annotation of the MHV68 transcriptome.
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Gammaherpesvirinae/metabolismo , Infecciones por Herpesviridae/metabolismo , Sistemas de Lectura Abierta , ARN Viral/biosíntesis , Transcriptoma , Animales , Estudio de Asociación del Genoma Completo , Ratones , Células 3T3 NIHAsunto(s)
Carbunco/diagnóstico , Enfermedades Cutáneas Bacterianas/diagnóstico , Adulto , Animales , Carbunco/diagnóstico por imagen , Carbunco/tratamiento farmacológico , Antibacterianos/uso terapéutico , Bovinos , Brotes de Enfermedades/prevención & control , Femenino , Fiebre/tratamiento farmacológico , Fiebre/etiología , Humanos , Linfangitis/etiología , Carne/microbiología , Persona de Mediana Edad , Penicilina G/uso terapéutico , Enfermedades Cutáneas Bacterianas/diagnóstico por imagen , Enfermedades Cutáneas Bacterianas/tratamiento farmacológico , Turquía/epidemiologíaRESUMEN
Gammaherpesviruses, including the human pathogens Epsteinâ»Barr virus (EBV) and Kaposi's sarcoma-associated herpesvirus (KSHV) are oncogenic viruses that establish lifelong infections in hosts and are associated with the development of lymphoproliferative diseases and lymphomas. Recent studies have shown that the majority of the mammalian genome is transcribed and gives rise to numerous long non-coding RNAs (lncRNAs). Likewise, the large double-stranded DNA virus genomes of herpesviruses undergo pervasive transcription, including the expression of many as yet uncharacterized lncRNAs. Murine gammaperherpesvirus 68 (MHV68, MuHV-4, ï§HV68) is a natural pathogen of rodents, and is genetically and pathogenically related to EBV and KSHV, providing a highly tractable model for studies of gammaherpesvirus biology and pathogenesis. Through the integrated use of parallel data sets from multiple sequencing platforms, we previously resolved transcripts throughout the MHV68 genome, including at least 144 novel transcript isoforms. Here, we sought to molecularly validate novel transcripts identified within the M3/M2 locus, which harbors genes that code for the chemokine binding protein M3, the latency B cell signaling protein M2, and 10 microRNAs (miRNAs). Using strand-specific northern blots, we validated the presence of M3-04, a 3.91 kb polyadenylated transcript that initiates at the M3 transcription start site and reads through the M3 open reading frame (ORF), the M3 poly(a) signal sequence, and the M2 ORF. This unexpected transcript was solely localized to the nucleus, strongly suggesting that it is not translated and instead may function as a lncRNA. Use of an MHV68 mutant lacking two M3-04-antisense pre-miRNA stem loops resulted in highly increased expression of M3-04 and increased virus replication in the lungs of infected mice, demonstrating a key role for these RNAs in regulation of lytic infection. Together these findings suggest the possibility of a tripartite regulatory relationship between the lncRNA M3-04, antisense miRNAs, and the latency gene M2.